Peng Liang-Hong, Shen Wei, Yong Wang, Lu Lu, Liu Lin
Department of Ophthalmology, General Hospital of Guangzhou Military Command of PLA, Guangzhou 510010, Guangdong Province, China.
Int J Ophthalmol. 2011;4(1):44-8. doi: 10.3980/j.issn.2222-3959.2011.01.10. Epub 2011 Feb 18.
To investigate the therapeutic effects of local and systemic administration of AMD3100 for alkali burn induced corneal neovascularization (CNV) in mice.
CNV was induced in vivo by alkaline burn of cornea in C57BL/6 mice. AMD3100 was administrated topically by subconjunctival injection or systemically by intraperitoneal injection for 7 days; balanced salt solution was administrated topically or systemically as a control respectively. Inflammatory index was evaluated by slit-lamp biomicroscopy and inflammatory cells infiltrated to cornea tissue were detected by histologic analysis at multiple time points. CNV was compared between the local and systemic treated mice 2 weeks after alkali burn, as quantified by CD34 immunostaining. Fluorescence-Activated Cell Sorter Analysis was used to investigate the mobilizing effects of EPC in mice after subconjunctival injected or intraperitoneal injected AMD3100. Immunohistochemistry was used to detect the expression of endothelial progenitor cells (EPC) marker proteins VEGFR2 and CD34.
Three days after alkali burn, infiltration of inflammatory cells was found in corneal tissue. At the first 7 days of local injection group, the number of inflammatory cells was significantly lower than that in systemic injection group. CNV could be seen at the 7(th) day, and at the 14(th) day reached the peak, then started to decrease. The number of CNV in the subconjunctival injection group was 7.57±1.26 per 0.034mm(2), compared to a number of 14.87±2.21 per 0.034mm(2) in the control group (P<0.05). On the contrary, the number of CNV in the intraperitoneal injection group was a little higher than that in the control group, 16.34±1.53 per 0.034mm(2)vs 13.26±1.87 per 0.034mm(2). The research also showed that intraperitoneally, but not subconjunctivally injected AMD3100 could mobilize EPC. On the other hand, subconjunctival, but not intraperitoneally injected AMD3100 could reduce the expression of EPC marker proteins.
In mice locally administrated AMD3100 can reduce the number of alkali burn induced CNV. The number of inflammatory cells and inflammatory responses in corneal tissue.
研究局部和全身应用AMD3100对小鼠碱烧伤诱导的角膜新生血管化(CNV)的治疗效果。
通过对C57BL/6小鼠角膜进行碱烧伤在体内诱导CNV。AMD3100通过结膜下注射局部给药或通过腹腔注射全身给药7天;分别以平衡盐溶液局部或全身给药作为对照。在多个时间点通过裂隙灯生物显微镜评估炎症指数,并通过组织学分析检测浸润到角膜组织中的炎症细胞。在碱烧伤2周后比较局部和全身治疗小鼠的CNV,通过CD34免疫染色进行定量。荧光激活细胞分选分析用于研究结膜下注射或腹腔注射AMD3100后小鼠中内皮祖细胞(EPC)的动员作用。免疫组织化学用于检测内皮祖细胞(EPC)标志物蛋白VEGFR2和CD34的表达。
碱烧伤后3天,在角膜组织中发现炎症细胞浸润。在局部注射组的前7天,炎症细胞数量明显低于全身注射组。在第7天可见CNV,在第14天达到峰值,然后开始下降。结膜下注射组每0.034mm²的CNV数量为7.57±1.26,而对照组为每0.034mm² 14.87±2.21(P<0.05)。相反,腹腔注射组每0.034mm²的CNV数量略高于对照组,为16.34±1.53 vs 13.26±1.87每0.034mm²。研究还表明,腹腔注射而非结膜下注射AMD3100可动员EPC。另一方面,结膜下注射而非腹腔注射AMD3100可降低EPC标志物蛋白的表达。
在小鼠中局部应用AMD3100可减少碱烧伤诱导的CNV数量、角膜组织中的炎症细胞数量和炎症反应。